Table 4.
Robustness Testinga
| method parameters | current method parameter | result at current level (nmol/g Hb) | parameter at lower level | result at lower level (nmol/g Hb) | parameter at higher level | result at higher level (nmol/g Hb) |
|---|---|---|---|---|---|---|
| pH of digest buffer | 8.5 | 51.1 ± 3.2 | 8.0 | 50.6 ± 4.9 | 9.0 | 49.9 ± 2.7 |
| concentration of digest buffer (mM) | 200 | 51.1 ± 3.2 | 150 | 52.3 ± 2.9 | 250 | 45.1 ± 1.6b |
| digestion time (h) | 48 | 51.5 ± 3.3 | 44 | 49.2 ± 2.9 | 50 | 51.0 ± 1.6 |
| amount of trypsin (μg) | 16.7 | 43.0 ± 3.7 | 12.5 | 40.3 ± 2.5 | 23.5 | 45.6 ± 3.4 |
| amount of hemoglobin (mg) | 500 | 38.9 ± 4.4 | 425 | 36.5 ± 2.7 | 575 | 37.5 ± 2.9 |
a
FA-VHLTPEEK concentrations values at sample preparation conditions above and below those used in the method.
b
Significantly different from values obtained with current method parameter, p < 0.05 using two-way ANOVA model.